Mitogen-activated protein kinase translocates into the germinal vesicle and induces germinal vesicle breakdown in porcine oocytes.
The role of mitogen-activated protein kinase (MAPK) in the meiotic resumption of porcine oocytes was examined. First, using indirect immunofluorescence staining with a specific antibody against rat MAPK, we monitored the dynamics of the subcellular distribution of MAPK during meiosis initiation. We found that the inactive MAPK was already present in immature oocytes arrested at the G2 stage and that this inactive kinase was localized exclusively in the cytoplasm. At the G2/M transition stage, part of the MAPK moved into the germinal vesicle (GV) before germinal vesicle breakdown (GVBD). In addition, immunoblot analysis showed that the nuclear MAPK existed in an active form. To determine whether this active MAPK could induce GVBD, we microinjected active MAPK into immature porcine oocytes. The active MAPK injected into the cytoplasm was quickly inactivated and could not accelerate GVBD. In contrast, MAPK injection into the GV markedly accelerated GVBD. These results show that in porcine oocytes, 1) inactive MAPK localizes in the cytosol of immature GV oocytes, 2) part of the activated MAPK translocates into the GV just before GVBD, and 3) exogenous MAPK maintains its activity level in the GV and induces GVBD, indicating that MAPK mediates the maturation-inducing signal from the cytoplasm into the nucleus and induces meiosis reinitiation.
Inoue
M
M
Department of Reproductive and Developmental Biology, The Institute of Medical Science, The University of Tokyo, Japan.
Naito
K
K
Nakayama
T
T
Sato
E
E
D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000008:administration & dosage / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005455:Fluorescent Antibody Technique; D008540:Meiosis; D008845:Microinjections; D009865:Oocytes / Q000201:enzymology* / Q000648:ultrastructure; D013552:Swine
D017871:Calcium-Calmodulin-Dependent Protein Kinases